1. Chapter 9 Nuclear Magnetic Resonance and Mass Spectrometry

2. Introduction • Classic methods for organic structure determination: • Boiling point • Refractive index • Solubility tests • Functional group tests • Derivative preparation • Sodium fusion (to identify N, Cl, Br, I & S) • Mixture melting point • Combustion analysis • Degradation

3. Classic methods for organic structure determination. • These methods require large quantities of sample and are time consuming.

4. Spectroscopic methods for organic structure determination. • Mass Spectroscopy (MS) • Molecular Mass & characteristic fragmentationpattern • Infrared Spectroscopy (IR) • Characteristic functional groups • Ultraviolet Spectroscopy (UV) • Characteristic chromophore • Nuclear Magnetic Resonance (NMR)

5. Spectroscopic methods for organic structure determination. • Combination of these spectroscopic techniques provides a rapid, accurate and powerful tool for Identification and Structure Elucidation of organic compounds. • Effective in mg and microgram quantities.

6. General steps for structure elucidation. • Elemental analysis • Empirical formula • e.g. C2H4O • Mass spectroscopy • Molecular weight • Molecular formula • e.g. C4H8O2, C6H12O3 … etc. • Characteristic fragmentation pattern for certain functional groups.

7. General steps for structure elucidation. • From molecular formula • Double bond equivalent (DBE) • Infrared spectroscopy (IR) • Identify some specific functional groups • e.g. C=O, C–O, O–H, COOH, NH2 … etc.

8. General steps for structure elucidation. • UV • Sometimes useful especially for conjugated systems. • e.g. dienes, aromatics, enones • 1H, 13C NMR and other advanced NMR techniques. • Full structure determination

9. Electromagnetic spectrum

10. Nuclear Magnetic Resonance(NMR) Spectroscopy • A graph that shows the characteristic energy absorption frequencies and intensities for a sample in a magnetic field is called a nuclear magnetic resonance (NMR) spectrum.

11. Typical 1H (Proton) NMR spectrum

12. The number of signals in the spectrum tells us how many different sets of protons there are in the molecule. The position of the signals in the spectrum along the x-axis tells us about the magnetic environment of each set of protons arising largely from the electron density in their environment.

13. The area under the signal tells us about how many protons there are in the set being measured. The multiplicity (or splitting pattern) of each signal tells us about the number of protons on atoms adjacent to the one whose signal is being measured.

14. Typical 1H NMR spectrum • Chemical Shift (). • Integration (areas of peaks  no. of H). • Multiplicity (spin-spin splitting) and coupling constant.

15. Typical 1H NMR spectrum

16. 2A. Chemical Shift • The position of a signal along the x-axis of an NMR spectrum is called its chemical shift. • The chemical shift of each signal gives information about the structural environment of the nuclei producing that signal. • Counting the number of signals in a 1H NMR spectrum indicates, at a first approximation, the number of distinct proton environments in a molecule.

17. 1H Chemical Shifts

18. 13C Chemical Shifts

19. Normal range of 1H NMR

20. Reference compound: • TMS = tetramethylsilane as a reference standard (0 ppm). • Reasons for the choice of TMS: • Resonance position at higher field than other organic compounds. • Unreactive and stable, not toxic • Volatile and easily removed (B.P. = 28oC)

21. NMR solvent • Normal NMR solvents should not contain hydrogen. • Common solvents: • CDCl3 • C6D6 • CD3OD • CD3COCD3 (d6-acetone)

22. The 300-MHz 1H NMR spectrum of 1,4-dimethylbenzene. • Peaks are in blue, integration in black.

23. 2B. Integration of Signal Areas Integral Step Heights • The area under each signal in a 1H NMR spectrum is proportional to the number of hydrogen atoms producing that signal. • It is signal area (integration), not signal height, that gives information about the number of hydrogen atoms.

24. 3 Hb 2 Ha Splitting of protons on adjacent atoms. Hb Ha

25. 2C. Coupling (Signal Splitting) • Coupling is caused by the magnetic effect of nonequivalent hydrogen atoms that are within 2 or 3 bonds of the hydrogens producing the signal. • The n+1 rule • Rule of Multiplicity: If a proton (or a set of magnetically equivalent nuclei) has n neighbors of magnetically equivalent protons. It’s multiplicity is n + 1.

26. Examples

27. Example of splitting

28. Examples Note: All Hb’s are chemically and magnetically equivalent.

29. Pascal’s Triangle: • Used to predict relative intensity of various peaks in multiplet. • Given by the coefficient of binomial expansion (a + b)n. singlet (s) 1 doublet (d) 1 1 triplet (t) 1 2 1 quartet (q) 1 3 3 1 quintet 1 4 6 4 1 sextet 1 5 10 10 5 1

30. Pascal’s Triangle • For • For • Due to symmetry, Ha and Hb are identical •  a singlet • Ha ≠ Hb •  two doublets

31. How to Interpret Proton NMRSpectra Count the number of signals to determine how many distinct proton environments are in the molecule (neglecting, for the time being, the possibility of overlapping signals). Use chemical shift tables or charts to correlate chemical shifts with possible structural environments.

32. Determine the relative area of each signal, as compared with the area of other signals, as an indication of the relative number of protons producing the signal. Interpret the splitting pattern for each signal to determine how many hydrogen atoms are present on carbon atoms adjacent to those producing the signal and sketch possible molecular fragments. Join the fragments to make a molecule in a fashion that is consistent with the data.

33. Example: 1H NMR (300 MHz) of an unknown compound with molecular formula C3H7Br.

34. Three distinct signals at ~ d3.4, 1.8 and 1.1 ppm.  d3.4 ppm: likely to be near an electronegative group (Br).

35. d (ppm): 3.41.81.1 Integral: 223

36. d (ppm): 3.41.81.1 Multiplicity: tripletsextettriplet 2 H's on adjacent C 5 H's on adjacent C 2 H's on adjacent C

37. Complete structure: most upfield signal most downfield signal • 2 H's from integration • triplet • 2 H's from integration • sextet • 3 H's from integration • triplet

38. Nuclear Spin:The Origin of the Signal The magnetic field associated with a spinning proton The spinning proton resembles a tiny bar magnet

39. Alignment of nuclei in presence and absence of an applied magnetic field: random with or against

40. Difference in energy is proportional to applied field strength.

41. Spin quantum number (I) 1H: I = ½ (two spin states: +½ or -½)  (similar for 13C, 19F, 31P) 12C, 16O, 32S: I = 0  These nuclei do not give an NMR spectrum

42. Detecting the Signal: Fourier Transform NMR Spectrometers

44. Shielding & Deshielding of Protons • All protons do not absorb energy at the same frequency in a given external magnetic field. • Lower chemical shift values correspond with lower frequency. • Higher chemical shift values correspond with higher frequency.

46. Deshielding by electronegative groups • Greater electronegativity • Deshielding of the proton • Larger d

47. Shielding and deshielding by circulation of p electrons: • If we were to consider only the relative electronegativities of carbon in its three hybridization states, we might expect the following order of protons attached to each type of carbon: (higher frequency) (lower frequency) sp < sp2 < sp3

48. In fact, protons of terminal alkynes absorb between d 2.0 and d 3.0, and the order is: (higher frequency) (lower frequency) sp2 < sp < sp3

49. This upfield shift (lower frequency) of the absorption of protons of terminal alkynes is a result of shielding produced by the circulating p electrons of the triple bond (anisotropy). Shielded (d 2 – 3 ppm)

50. Shielded region Deshielded region • Aromatic system